LOCUS       Exported                6240 bp ds-DNA     circular SYN 16-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6240)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 20, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6240
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..2953
                     /organism="Homo sapiens"
                     /chromosome="14"
                     /map="14q32.32"
                     /mol_type="mRNA"
                     /db_xref="taxon:9606"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     primer_bind     141..190
                     /label=F-D1
     primer_bind     147..194
                     /label=F-Mlu-D2
     primer_bind     147..161
                     /label=3'-pcalm-ampF
     primer_bind     150..168
                     /label=P-Calm1-F
     primer_bind     151..200
                     /label=F-D2
     misc_feature    156..276
                     /label=P Calm1
     misc_feature    156..165
                     /label=D1
     primer_bind     161..210
                     /label=F-D3
     primer_bind     complement(161..185)
                     /label=R-D4
     misc_feature    166..175
                     /label=D2
     primer_bind     171..220
                     /label=F-D4
     primer_bind     complement(171..195)
                     /label=R-D5
     misc_feature    176..185
                     /label=D3
     primer_bind     181..230
                     /label=F-D5
     primer_bind     complement(181..205)
                     /label=R-D6
     misc_feature    186..195
                     /label=D4
     primer_bind     191..240
                     /label=F-D6
     primer_bind     complement(191..215)
                     /label=r-d7
     misc_feature    196..205
                     /label=D5
     primer_bind     200..250
                     /label=F-D7
     primer_bind     complement(201..225)
                     /label=R-D8
     misc_feature    206..215
                     /label=D6
     primer_bind     211..260
                     /label=F-D8
     primer_bind     complement(215..236)
                     /label=R-D9
     misc_feature    216..225
                     /label=D7
     primer_bind     222..271
                     /label=F-D9
     primer_bind     complement(222..246)
                     /label=R-D10
     misc_feature    226..235
                     /label=D8
     primer_bind     complement(232..256)
                     /label=R-D11
     misc_feature    236
                     /label=+1
     misc_feature    237..246
                     /label=D9
     primer_bind     complement(242..266)
                     /label=R-D12
     misc_feature    247..256
                     /label=D10
     misc_feature    257..266
                     /label=D11
     primer_bind     complement(260..287)
                     /label=2B-1R
     primer_bind     complement(261..276)
                     /label=P-Calm1-R
     misc_feature    267..276
                     /label=D12
     primer_bind     271..296
                     /label=2B-2F
     primer_bind     complement(277..298)
                     /label=Primer 1
     misc_feature    277..282
                     /label=BsiWI
     primer_bind     283..302
                     /label=Primer 5
     CDS             288..1318
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    548
                     /label=silent mut
     primer_bind     complement(616..650)
                     /label=R-Cre-R119STOP
     primer_bind     complement(632..651)
                     /label=R-CRE-L115-BamHI
     primer_bind     636..670
                     /label=F-Cre-R119STOP
     primer_bind     1093..1116
                     /label=F-Seq-Cre-3F1
     misc_feature    1093..1098
                     /label=ClaI
     misc_feature    1224
                     /label=mut
     primer_bind     1262..1279
                     /label=cre-SF
     primer_bind     complement(1296..1318)
                     /label=R-T5-Cre
     intron          1320..1360
                     /label=Splicing donor
                     /note="Splicing donor"
     primer_bind     complement(1338..1367)
                     /label=2B-2R
     primer_bind     complement(1341..1385)
                     /label=Primer 7
     primer_bind     complement(1341..1385)
                     /label=Primer 6
     primer_bind     1350..1387
                     /label=2B-3F
     primer_bind     complement(1356..1380)
                     /label=Primer 3
     primer_bind     1367..1401
                     /label=Primer 8
     primer_bind     1367..1401
                     /label=Primer 2
     primer_bind     1367..1401
                     /label=F-T5-JT+2272
     protein_bind    1367..1400
                     /label=loxJTZ17?2272?
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1380..1387
                     /label=2272
     intron          1401..1492
                     /label=Splicing acceptor
                     /note="Splicing acceptor"
     primer_bind     1401..1420
                     /label=Primer 4
     primer_bind     1459..1478
                     /label=T0060_PIRES-F
     primer_bind     complement(1468..1492)
                     /label=#57-R-SA-3R1
     primer_bind     1477..1511
                     /label=B47F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..2953
                     /codon_start=1
                     /gene="CDC42BPB"
                     /gene_synonym="CHOCNS; MRCKB"
                     /product="serine/threonine-protein kinase MRCK beta
                     isoform 1"
                     /EC_number="2.7.11.1"
                     /label=CDC42BPB
                     /note="isoform 1 is encoded by transcript variant 1; 
                     serine/threonine-protein kinase MRCK beta; myotonic 
                     dystrophy protein kinase-like beta; DMPK-like beta; 
                     myotonic dystrophy kinase-related CDC42-binding kinase 
                     beta; CDC42BP-beta; CDC42 binding protein kinase beta 
                     (DMPK-like)"
                     /db_xref="GI:115527097"
                     /db_xref="CCDS:CCDS9978.1"
                     /db_xref="GeneID:9578"
                     /db_xref="HGNC:HGNC:1738"
                     /db_xref="MIM:614062"
                     /protein_id="NP_006026.3"
                     /translation="VVHVPLEAYDSSLPLIKAILTAAIVDADRIAVGLEEGLYVIEVTR
                     DVIVRAADCKKVHQIELAPREKIVILLCGRNHHVHLYPWSSLDGAEGSFDIKLPETKGC
                     QLMATATLKRNSGTCLFVAVKRLILCYEIQRTKPFHRKFNEIVAPGSVQCLAVLRDRLC
                     VGYPSGFCLLSIQGDGQPLNLVNPNDPSLAFLSQQSFDALCAVELESEEYLLCFSHMGL
                     YVDPQGRRARAQELMWPAAPVACSCSPTHVTVYSEYGVDVFDVRTMEWVQTIGLRRIRP
                     LNSEGTLNLLNCEPPRLIYFKSKFSGAVLNVPDTSDNSKKQMLRTRSKRRFVFKVPEEE
                     RLQQRREMLRDPELRSKMISNPTNFNHVAHMGPGDGMQVLMDLPLSAVPPSQEERPGPA
                     PTNLARQPPSRNKPYISWPSSGGSEPSVTVPLRSMSDPDQDFDKEPDSDSTKHSTPSNS
                     SNPSGPPSPNSPHRSQLPLEGLEQPACDT"
     misc_feature    1493..1494
                     /label=Feature 58
     primer_bind     complement(2933..2974)
                     /label=Primer 1
     primer_bind     complement(2933..2974)
                     /label=nB47F-BsrGI-R
     primer_bind     complement(2933..2974)
                     /label=B47F-BsrGI-R
     misc_feature    2954..2959
                     /label=BsrGI
     CDS             2960..3025
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2965..3053)
                     /label=296
     terminator      3029..3460
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3029..3054
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3108..3131)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3503..3643
                     /label=R-ITR
     rep_origin      3646..4101
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3793..3802)
                     /label=F-Seq-Cre-3F1
     promoter        4383..4487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4488..5348
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="HSNMYPLMRQ*P**MLQ*Y*KRKSMSIQHFRVALIPFFAAFCLPV
                     FAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLS
                     RIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTT
                     IGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLT
                     LASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIV
                     VIYTTG"
     rep_origin      5519..6107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcgctc tgcgcgaggc ggctccgccg
      181 cggcggaggg atacggcgca ccatatatat atcgcggggc gcagactcgc gctccggcag
      241 tggtgctggg agtgtcgtgg acgccgtgcc gttactcgta cgccaccatg tccaatttac
      301 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      361 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      421 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      481 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      541 taaaaaccat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      601 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      661 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      721 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      781 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      841 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      901 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      961 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
     1021 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1081 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1141 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1201 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgta atgaactata
     1261 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagg
     1321 taagtatcaa ggttacaaga caggtttaag gagaccaata actagtataa cttcgtatag
     1381 gatactttat agcaatttat gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1441 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtcgtgca
     1501 tgttcccttg gaagcctacg acagctcgct gcctctcatc aaggccatcc tgacagctgc
     1561 catcgtggat gcagacagga ttgcagtcgg cctagaagaa gggctctatg tcatagaggt
     1621 cacccgagat gtgatcgtcc gtgccgctga ctgtaagaag gtacaccaga tcgagcttgc
     1681 tcccagggag aagatcgtaa tcctcctctg tggccggaac caccatgtgc acctctatcc
     1741 gtggtcgtcc cttgatggag cggaaggcag ctttgacatc aagcttccgg aaaccaaagg
     1801 ctgccagctc atggccacgg ccacactcaa gaggaactct ggcacctgcc tgtttgtggc
     1861 cgtgaaacgg ctgatccttt gctatgagat ccagagaacg aagccattcc acagaaagtt
     1921 caatgagatt gtggctcccg gcagcgtgca gtgcctggcg gtgctcaggg acaggctctg
     1981 tgtgggctac ccttctgggt tctgcctgct gagcatccag ggggacgggc agcctctaaa
     2041 cctggtaaat cccaatgacc cctcgcttgc gttcctctca caacagtctt ttgatgccct
     2101 ttgtgctgtg gagctcgaaa gcgaggagta cctgctttgc ttcagccaca tgggactgta
     2161 cgtggacccg caaggccgga gggcacgcgc gcaggagctc atgtggcctg cggctcctgt
     2221 cgcctgtagt tgcagcccca cccacgtcac ggtgtacagc gagtatggcg tggacgtctt
     2281 tgatgtgcgc accatggagt gggtgcagac catcggcctg cggaggataa ggcccctgaa
     2341 ctctgaaggc accctcaacc tcctcaactg cgagcctcca cgcttgatct acttcaagag
     2401 caagttctcg ggagcggttc tcaacgtgcc ggacacctcc gacaacagca agaagcagat
     2461 gctgcgcacc aggagcaaaa ggcggttcgt cttcaaggtc ccagaggaag agagactgca
     2521 gcagaggcga gagatgctta gagacccaga attgagatcc aaaatgatat ccaacccaac
     2581 caacttcaac cacgtggccc acatgggccc aggcgacggc atgcaggtgc tcatggacct
     2641 gcctctgagt gctgtgcccc cctcccagga ggaaaggccg ggccccgctc ccaccaacct
     2701 ggctcgccag cctccatcca ggaacaagcc ctacatctcg tggccctcat caggtggatc
     2761 ggagcctagc gtgactgtgc ctctgagaag tatgtctgat ccagaccagg actttgacaa
     2821 agagcctgat tcggactcca ccaaacactc aactccatcg aatagctcca accccagcgg
     2881 cccaccgagc cccaactccc cccacaggag ccagctcccc ctcgaaggcc tggagcagcc
     2941 ggcctgtgac acctgtacag actacaaaga ccatgacggt gattataaag atcatgacat
     3001 cgactacaag gatgacgatg acaagtaact cgagataatc aacctctgga ttacaaaatt
     3061 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3121 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3181 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt
     3301 gatgctattg ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc
     3361 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3421 tatgtttcag gttcaggggg agatgtggga ggttttttaa agatctggag accggtgtgg
     3481 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3541 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3601 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3661 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3721 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3781 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3841 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3901 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3961 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4021 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4081 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4141 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4201 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4261 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4321 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4381 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4441 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4501 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4561 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4621 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4681 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4741 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4801 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4861 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4921 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4981 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5041 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5101 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5161 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5221 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5281 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5341 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5401 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5701 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
